We are dealing with culturable analysis of microbial community found in hot springs. Which procedures should I apply for studying cultivable microbial diversity and how can we start our work in the laboratory? Provide some basic idea for these work like media, parameters etc.
Wow! This is a very broad based question.
However, the first thing you would want to ascertain is the temperature of the hot springs from which you wish to study the microflora.
It may be as low as 70C or as high as 90C or even higher.
To start off take perhaps five samples (1 ml each) in Eppedorf tubes and microfuge them to get a pellet of the organisms in your sample.
Resuspend each pellet in about 0,2 ml of phoasphate buffered saline, make a smear on a slide of each and do a Gram's staining. This will give you an idea of the morphology of the kinds of organisms present in your sample.
Combine the remaining pellets and add 5 ml of a nutrient medium such as Trypticase Soy Broth or Nutrient broth. Incubate the tube at the temperature closes to that of the hot spring from which the original sample was obtained.
Check every 24 hours for turbidity (growth) and do a Gram's stain of any growth seen.
The above should give you a good starting point. We can discuss further via email if you wish.
Good luck. God bless.
Wow! This is a very broad based question.
However, the first thing you would want to ascertain is the temperature of the hot springs from which you wish to study the microflora.
It may be as low as 70C or as high as 90C or even higher.
To start off take perhaps five samples (1 ml each) in Eppedorf tubes and microfuge them to get a pellet of the organisms in your sample.
Resuspend each pellet in about 0,2 ml of phoasphate buffered saline, make a smear on a slide of each and do a Gram's staining. This will give you an idea of the morphology of the kinds of organisms present in your sample.
Combine the remaining pellets and add 5 ml of a nutrient medium such as Trypticase Soy Broth or Nutrient broth. Incubate the tube at the temperature closes to that of the hot spring from which the original sample was obtained.
Check every 24 hours for turbidity (growth) and do a Gram's stain of any growth seen.
The above should give you a good starting point. We can discuss further via email if you wish.
Good luck. God bless.
Hi,
just to add something to the answer above. Concerning the required temperatures, evaporation is realy nasty problem. In similar experiments for estimation of the number of bacteria in termal spring (70C), I used plates and partly overcome the problem with parafilm.
Wish You luck
my first suggestion is to search for "thermophilic & sp. nov." in Pubmed. There are plenty of papers describing the isolation of novel thermophilic m-o. In general, the majority of thermophiles are anaerobes, thus you need to prepare anaerobic medium and use sealed tubes (in this case evaporation is not a problem). Also, you need to inoculate these tubes with your sample material ASAP avoiding any preliminary manipulations (like Gram staining etc. actually, Gram staining is being completely obsolete method) or perform these manipulations under sterile nitrogen or argon or CO2, whatever. Anyway, search details in papers
Исследователи, выделяющие микробы из термальных вод, встречали ли вы стафилококки?
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